Related Experiment Video
Updated: Jan 19, 2026

In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 Ehmeth Enzyme
Published on: October 19, 2010
Yeast PAF1 complex counters the pol III accumulation and replication stress on the tRNA genes
Pratibha Bhalla1,2, Ashutosh Shukla1,2, Dipti Vinayak Vernekar1,3
1Centre for Cellular and Molecular Biology (Council of Scientific and Industrial Research), Hyderabad, India.
Abstract:
The RNA polymerase (pol) III transcribes mostly short, house-keeping genes, which produce stable, non-coding RNAs. The tRNAs genes, highly transcribed by pol III in vivo are known replication fork barriers. One of the transcription factors, the PAF1C (RNA polymerase II associated factor 1 complex) is reported to associate with pol I and pol II and influence their transcription. We found low level PAF1C occupancy on the yeast pol III-transcribed genes, which is not correlated with nucleosome positions, pol III occupancy and transcription. PAF1C interacts with the pol III transcription complex and causes pol III loss from the genes under replication stress. Genotoxin exposure causes pol III but not Paf1 loss from the genes. In comparison, Paf1 deletion leads to increased occupancy of pol III, γ-H2A and DNA pol2 in gene-specific manner. Paf1 restricts the accumulation of pol III by influencing the pol III pause on the genes, which reduces the pol III barrier to the replication fork progression.
Related Concept Videos
12:36In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
10:41Continuous High-resolution Microscopic Observation of Replicative Aging in Budding Yeast
12:41Measuring Replicative Life Span in the Budding Yeast
tRNA Activation
09:53Mitochondrial Transformation in Baker's Yeast to Study Translation and Respiratory Complex Assembly
10:34Using an Automated Cell Counter to Simplify Gene Expression Studies: siRNA Knockdown of IL-4 Dependent Gene Expression in Namalwa Cells

